LOCUS       sequence-216909-        7703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BTK Type 8 Bacterial origin (broad-host-range) for golden gate 
            assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7703)
  AUTHORS   Leonard SP, Perutka J, Powell JE, Geng P, Richhart DD, Byrom M, Kar 
            S, Davies BW, Ellington AD, Moran NA, Barrick JE
  TITLE     Genetic Engineering of Bee Gut Microbiome Bacteria with a Toolkit 
            for Modular Assembly of Broad-Host-Range Plasmids.
  JOURNAL   ACS Synth Biol. 2018 May 18;7(5):1279-1290. doi: 
            10.1021/acssynbio.7b00399. Epub 2018 Apr 13.
  PUBMED    29608282
REFERENCE   2  (bases 1 to 7703)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.7b00399"; journalName: "ACS Synth Biol"; date: 
            "2018-05-18- 18"; volume: "7"; issue: "5"; pages: "1279-1290"
COMMENT     Sequence Label: pBTK401
FEATURES             Location/Qualifiers
     source          1..7703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      292..686
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     protein_bind    1138..1156
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1163..1181
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1199..1210
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1217..1894
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1912..1983
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1999..2026
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(2150..2941,2942..3010))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2773..2792
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3011..3115)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(3168..4019)
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(4006..4845)
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             complement(5356..6327)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     oriT            7566..7653
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
ORIGIN
        1 ggcggcggat cggctggcca agctcgaaga acaacgagcg cgaatcaatg ccgaaattca
       61 gcgggtgcgg gcaagggaac agcagcaaga gcgcaagaac gaaacaaggc gcaaggtgct
      121 ggtgggggcc atgattttgg ccaaggtgaa cagcagcgag tggccggagg atcggctcat
      181 ggcggcaatg gatgcgtacc ttgaacgcga ccacgaccgc gccttgttcg gtctgccgcc
      241 acgccagaag gatgagccgg gctgaatgat cgaccgagac aggccctgcg gggctgcaca
      301 cgcgccccca cccttcgggt agggggaaag gccgctaaag cggctaaaag cgctccagcg
      361 tatttctgcg gggtttggtg tggggtttag cgggctttgc ccgcctttcc ccctgccgcg
      421 cagcggtggg gcggtgtgta gcctagcgca gcgaatagac cagctatccg gcctctggcc
      481 gggcatattg ggcaagggca gcagcgcccc acaagggcgc tgataaccgc gcctagtgga
      541 ttattcttag ataatcatgg atggattttt ccaacacccc gccagccccc gcccctgctg
      601 ggtttgcagg tttgggggcg tgacagttat tgcaggggtt cgtgacagtt attgcagggg
      661 ggcgtgacag ttattgcagg ggttcgtgac agttagtacg ggagtgacgg gcactggctg
      721 gcaatgtcta gcaacggcag gcatttcggc tgagggtaaa agaactttcc gctaagcgat
      781 agactgtatg taaacacagt attgcaagga cgcggaacat gcctcatgtg gcggccagga
      841 cggccagccg ggatcgggat actggtcgtt accagagcca ccgacccgag caaacccttc
      901 tctatcagat cgttgacgag tattacccgg cattcgctgc gcttatggca gagcagggaa
      961 aggaattgcc gggctatgtg caacgggaat ttgaagaatt tctccaatgc gggcggctgg
     1021 agcatggctt tctacgggtt cgctgcgagt cttgccacgc cgagcacctg gtcgctttca
     1081 gaaatcaatc taaagtatat atgagtaaac ttggtctgac aggccccttg agaccagtcc
     1141 ctatcagtga tagagattga catccctatc agtgatagag atactgagca cggatctgaa
     1201 agaggagaaa ggatctatgg cgagtagcga agacgttatc aaagagttca tgcgtttcaa
     1261 agttcgtatg gaaggttccg ttaacggtca cgagttcgaa atcgaaggtg aaggtgaagg
     1321 tcgtccgtac gaaggtactc agaccgctaa actgaaagtt accaaaggtg gtccgctgcc
     1381 gttcgcttgg gacatcctgt ccccgcagtt ccagtacggt tccaaagctt acgttaaaca
     1441 cccggctgac atcccggact acctgaaact gtccttcccg gaaggtttca aatgggaacg
     1501 tgttatgaac ttcgaagacg gtggtgttgt taccgttacc caggactcct ccctgcaaga
     1561 cggtgagttc atctacaaag ttaaactgcg tggtactaac ttcccgtccg acggtccggt
     1621 tatgcagaaa aaaaccatgg gttgggaagc ttccaccgaa cgtatgtacc cggaagacgg
     1681 tgctctgaaa ggtgaaatca aaatgcgtct gaaactgaaa gacggtggtc actacgacgc
     1741 tgaagttaaa accacctaca tggctaaaaa accggttcag ctgccgggtg cttacaaaac
     1801 cgacatcaaa ctggacatca cctcccacaa cgaagactac accatcgttg aacagtacga
     1861 acgtgctgaa ggtcgtcact ccaccggtgc ttaataagga tctccaggca tcaaataaaa
     1921 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     1981 ctctactaga gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta taagtcggtc
     2041 tcaccgagcg gccgcgatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2101 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2161 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     2221 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     2281 tgataccgcg ggacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     2341 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     2401 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     2461 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     2521 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     2581 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     2641 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     2701 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     2761 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     2821 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     2881 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     2941 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3001 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3061 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3121 ttccccgaaa agtgccacct gtcatgacca aaatccctta acgtgagtca gcctgccgcc
     3181 ttgggccggg tgatgtcgta cttgcccgcc gcgaactcgg ttaccgtcca gcccagcgcg
     3241 accagctccg gcaacgcctc gcgcacccgc tggcggcgct tgcgcatggt cgaaccactg
     3301 gcctctgacg gccagacata gccgcacaag gtatctatgg aagccttgcc ggttttgccg
     3361 gggtcgatcc agccacacag ccgctggtgc agcaggcggg cggtttcgct gtccagcgcc
     3421 cgcacctcgt ccatgctgat gcgcacatgc tggccgccac ccatgacggc ctgcgcgatc
     3481 aaggggttca gggccacgta caggcgcccg tccgcctcgt cgctggcgta ctccgacagc
     3541 agccgaaacc cctgccgctt gcggccattc tgggcgatga tggatacctt ccaaaggcgc
     3601 tcgatgcagt cctgtatgtg cttgagcgcc ccaccactat cgacctctgc cccgatttcc
     3661 tttgccagcg cccgatagct acctttgacc accatggcat cagcggtgac ggcctcccac
     3721 ttgggttcca ggaacagccg gagctgccgt ccgccttcgg tcttgggttc cgggccaagc
     3781 actaggccat taggcccagc catggccacc agcccttgca ggatgcgcag atcatcagcg
     3841 cccagcggct ccgggccgct gaactcgatc cgcttgccgt cgccgtagtc atacgtcacg
     3901 tccagcttgc tgcgcttgcg ctcgccccgc ttgagggcac ggaacaggcc gggggccaga
     3961 cagtgcgccg ggtcgtgccg gacgtggctg aggctgtgct tgttcttagg cttcaccacg
     4021 gggcaccccc ttgctcttgc gctgcctctc cagcacggcg ggcttgagca ccccgccgtc
     4081 atgccgcctg aaccaccgat cagcgaacgg tgcgccatag ttggccttgc tcacaccgaa
     4141 gcggacgaag aaccggcgct ggtcgtcgtc cacaccccat tcctcggcct cggcgctggt
     4201 catgctcgac aggtaggact gccagcggat gttatcgacc agtaccgagc tgccccggct
     4261 ggcctgctgc tggtcgcctg cgcccatcat ggccgcgccc ttgctggcat ggtgcaggaa
     4321 cacgatagag cacccggtat cggcggcgat ggcctccatg cgaccgatga cctgggccat
     4381 ggggccgctg gcgttttctt cctcgatgtg gaaccggcgc agcgtgtcca gcaccatcag
     4441 gcggcggccc tcggcggcgc gcttgaggcc gtcgaaccac tccggggcca tgatgttggg
     4501 caggctgccg atcagcggct ggatcagcag gccgtcagcc acggcttgcc gttcctcggc
     4561 gctgaggtgc gccccaaggg cgtgcaggcg gtgatgaatg gcggtgggcg ggtcttcggc
     4621 gggcaggtag atcaccgggc cggtgggcag ttcgcccacc tccagcagat ccggcccgcc
     4681 tgcaatctgt gcggccagtt gcagggccag catggattta ccggcaccac cgggcgacac
     4741 cagcgccccg accgtaccgg ccaccatgtt gggcaaaacg tagtccagcg gtggcggcgc
     4801 tgctgcgaac gcctccagaa tattgatagg cttatgggta gccattgatt gcctcctttg
     4861 caggcagttg gtggttaggc gctggcgggg tcactacccc cgccctgcgc cgctctgagt
     4921 tcttccaggc actcgcgcag cgcctcgtat tcgtcgtcgg tcagccagaa cttgcgctga
     4981 cgcatccctt tggccttcat gcgctcggca tatcgcgctt ggcgtacagc gtcagggctg
     5041 gccagcaggt cgccggtctg cttgtccttt tggtctttca tatcagtcac cgagaaactt
     5101 gccggggccg aaaggcttgt cttcgcggaa caaggacaag gtgcagccgt caaggttaag
     5161 gctggccata tcagcgactg aaaagcggcc agcctcggcc ttgtttgacg tataaccaaa
     5221 gccaccgggc aaccaatagc ccttgtcact tttgatcagg tagaccgacc ctgaagcgct
     5281 tttttcgtat tccataaaac ccccttctgt gcgtgagtac tcatagtata acaggcgtga
     5341 gtaccaacgc aagcactaca tgctgaaatc tggcccgccc ctgtccatgc ctcgctggcg
     5401 gggtgccggt gcccgtgcca gctcggcccg cgcaagctgg acgctgggca gacccatgac
     5461 cttgctgacg gtgcgctcga tgtaatccgc ttcgtggccg ggcttgcgct ctgccagcgc
     5521 tgggctggcc tcggccatgg ccttgccgat ttcctcggca ctgcggcccc ggctggccag
     5581 cttctgcgcg gcgataaagt cgcacttgct gaggtcatca ccgaagcgct tgaccagccc
     5641 ggccatctcg ctgcggtact cgtccagcgc cgtgcgccgg tggcggctaa gctgccgctc
     5701 gggcagttcg aggctggcca gcctgcgggc cttctcctgc tgccgctggg cctgctcgat
     5761 ctgctggcca gcctgctgca ccagcgccgg gccagcggtg gcggtcttgc ccttggattc
     5821 acgcagcagc acccacggct gataaccggc gcgggtggtg tgcttgtcct tgcggttggt
     5881 gaagcccgcc aagcggccat agtggcggct gtcggcgctg gccgggtcgg cgtcgtactc
     5941 gctggccagc gtccgggcaa tctgcccccg aagttcaccg cctgcggcgt cggccacctt
     6001 gacccatgcc tgatagttct tcgggctggt ttccactacc agggcaggct cccggccctc
     6061 ggctttcatg tcatccaggt caaactcgct gaggtcgtcc accagcacca gaccatgccg
     6121 ctcctgctcg gcgggcctga tatacacgtc attgccctgg gcattcatcc gcttgagcca
     6181 tggcgtgttc tggagcactt cggcggctga ccattcccgg ttcatcatct ggccggtggt
     6241 ggcgtccctg acgccgatat cgaagcgctc acagcccatg gccttgagct gtcggcctat
     6301 ggcctgcaaa gtcctgtcgt tcttcatcgg gccaccaagc gcagccagat cgagccgtcc
     6361 tcggttgtca gtggcgtcag gtcgagcaag agcaacgatg cgatcagcag caccaccgta
     6421 ggcatcatgg aagccagcat cacggttagc catagcttcc agtgccaccc ccgcgacgcg
     6481 ctccgggcgc tctgcgcggc gctgctcacc tcggcggcta cctcccgcaa ctctttggcc
     6541 agctccaccc atgccgcccc tgtctggcgc tgggctttca gccactccgc cgcctgcgcc
     6601 tcgctggcct gctgggtctg gctcatgacc tgccgggctt cgtcggccag tgtcgccatg
     6661 ctctgggcca gcggttcgat ctgctccgct aactcgttga tgcctctgga tttcttcact
     6721 ctgtcgattg cgttcatggt ctattgcctc ccggtattcc tgtaagtcga tgatctgggc
     6781 gttggcggtg tcgatgttca gggccacgtc tgcccggtcg gtgcggatgc cccggccttc
     6841 catctccacc acgttcggcc ccaggtgaac accgggcagg cgctcgatgc cctgcgcctc
     6901 aagtgttctg tggtcaatgc gggcgtcgtg gccagcccgc tctaatgccc ggttggcatg
     6961 gtcggcccat gcctcgcggg tctgctcaag ccatgccttg ggcttgagcg cttcggtctt
     7021 ctgtgccccg cccttctccg gggtcttgcc gttgtaccgc ttgaaccact gagcggcggg
     7081 ccgctcgatg ccgtcattga tccgctcgga gatcatcagg tggcagtgcg ggttctcgcc
     7141 gccaccggca tggatggcca gcgtatacgg caggcgctcg gcaccggtca ggtgctgggc
     7201 gaactcggac gccagcgcct tctgctggtc gagggtcagc tcgaccggca gggcaaattc
     7261 gacctccttg aacagccgcc cattggcgcg ttcatacagg tcggcagcat cccagtagtc
     7321 ggcgggccgc tcgacgaact ccggcatgtg cccggattcg gcgtgcaaga cttcatccat
     7381 gtcgcgggca tacttgcctt cgcgctggat gtagtcggcc ttggccctgg ccgattggcc
     7441 gcccgacctg ctgccggttt tcgccgtaag gtgataaatc gccatgctgc ctcgctgttg
     7501 cttttgcttt tcggctccat gcaatggccc tcggagagcg caccgcccga agggtggccg
     7561 ttaggccagt ttctcgaaga gaaaccggta agtgcgccct cccctacaaa gtagggtcgg
     7621 gattgccgcc gctgtgcctc catgatagcc tacgagacag cacattaaca atggggtgtc
     7681 aagatggtta aggggagcaa caa
//
